Surfactants as a class of molecules are well-known to formulators of topically-applied products. Biosurfactants are a specific group of surfactants derived from naturally-occurring raw materials which can be easily degraded by proteases. Like standard surfactants they have both a water-soluble and a water-insoluble group on the same molecule, generally defined as a “head and tail”. As such, they have an affinity for both hydrophilic and lipophilic materials (e.g., oils, and more significantly for purposes of the present invention, cell membranes); thus, these are also described as amphipathic molecules. Biosurfactants of the present invention possess a high degree of affinity for cell membranes without the apparent disruption associated to standard surfactants at similar concentrations. They orientate themselves in a manner to lower surface tension between the incompatible “heads and tails”. As the concentration of biosurfactant increases, the interfacial surface becomes saturated, until a minimum surface tension, the so-called the critical micelle concentration (“CMC”), is reached. If biosurfactant is added beyond the CMC, micelles or aggregates form. The CMC is generally expressed in millimoles (mM) and is dependent on the temperature and ionic strength of the media. These aggregates vary in particle size and shape. The polymeric biosurfactant aggregates within the scope of the present invention typically have particle sizes in the nano-range, from about 5 to 100 nanometers.
In skin care products, sodium dodecyl sulfate is a commonly-used anionic surfactant that acts as a wetting agent, emulsifier or cleansing agent. It has a CMC in distilled water of about 8.13 mM (or ˜2400 ppm). Quaternary compounds are widely-used cationic surfactants. Dodecyl trimethyl ammonium bromide is representative of this class of compounds and has a CMC of about 14.6 mM (or ˜4300 ppm) in distilled water. By way of comparison, the CMC of phospholipids—the principal components of the cell membranes (e.g., diacyl phosphatidyl cholines)—range from about 5×10−3 mM to about 4.7×10−7 mM (˜3 ppm-˜0.003 ppm). See, e.g., D. Datta, Membrane Biochemistry (1987). Polymeric biosurfactants of the present invention have a CMC between the representative anionic and cationic compounds as well as cells membrane phospholipids discussed above.
Conventional surfactants, however, are known to cause irritation, inflammation and other negative sequelae. This is due, in part, to defatting the skin, removing necessary oils as well as rapid penetration to the epidermal layer. Surprisingly and unexpectedly, the polymeric biosurfactants of the present invention do not have these drawbacks at similar concentrations.
As discussed below, the use of amino acid sequences in skin care products is known in the art. Some such sequences are commercially available as acylated moieties (e.g., acetyl, myristoyl, palmitoyl). In general, acylation is a well-known technique to those of skill in the art for enhancing penetration of a water-loving or hydrophilic ingredient into the skin. The surface of normal skin is highly hydrophobic preventing significant penetration by hydrophilic substances. However, the properties of an acylated amino acid sequence can vary greatly in terms of toxicity which, in turn, affects its ultimate usefulness. Surprisingly and unexpectedly, many of the polymeric acylated biosurfactants of the present invention have comparatively low toxicity to mammalian cells (on the order of LD50>200) while at the same time maintaining a relatively high degree of toxicity for prokaryotic life forms.
Moreover, unlike prior art acylated amino acid sequences, the polymeric biosurfactants of the present invention have the ability to increase the synthesis of skin matrix proteins (e.g., elastin, fibronectin, collagen) and/or increase cell turnover rates while not causing a concomitant increase in the synthesis of enzymes that degrade these proteins (e.g., matrix metalloproteinases). Additionally, surprisingly and significantly, biosurfactants of the present invention do not cause an increase in inflammatory proteins, notably interleukin 6 and interleukin 8. This combination of properties makes these compounds uniquely suited to skin care applications.
The ability of polymeric biosurfactants of the present invention to effectively wet surfaces at low CMCs confers another surprising and unexpected property—broad spectrum antimicrobial activity. Polymeric biosurfactants of the present invention have the ability to inhibit the growth or kill a variety of microorganisms, including Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Staphylococcus aureus (S. aureus) and Candida albicans (C. albicans).
In the fields of personal care and dermatology, there has been, and continues to be, a need for highly effective multifunctional ingredients. The use of ingredients of this type helps to mitigate problems common to topical skin care formulations—e.g., instability (due to incompatibility of ingredients) as well decreased efficacy of active ingredients over time (due to interactions among the ingredients). Multifunctional ingredients—particularly those with low propensity to cause irritation, inflammation or other negative sequelae—are in high demand not only among formulators but also among increasingly demanding and sophisticated consumers. Taken in combination, the favorable properties of the polymeric biosurfactants of the present invention make them multifunctional ingredients that surprisingly and unexpectedly meet the heretofore unmet need for products with comparatively low toxicity that both help to restore, maintain and improve dermatologic conditions associated with disease, aging and/or environmental stressors while, in many instances, at the same time inhibiting microbial growth.
Prior Art Amino Acid Sequences Used in Skin Care Products
The following cosmetic ingredients, each consisting of two amino acids, are commercially available: Dipeptide-1 (Tyrosine and Arginine residues); Dipeptide-2 (Valine and Tryptophan residues); Dipeptide-4 (Phenylalanine and Tryptophan residues). Unless otherwise indicated, cosmetic ingredients are described by their assigned name in the International Cosmetic Ingredient (INCI) Dictionary and Handbook (10th Edition) published by the Cosmetic Toiletry and Fragrance Association (“CTFA”). The INCI Dictionary does not specify amino acid sequences or the amounts of each amino acid residue. As discussed below, a tripeptide may be described in the INCI Dictionary as containing two amino acid residues without indicating whether one of the two listed amino acids is present twice or whether the third amino acid in the peptide sequence is selected from the group of eighteen other naturally-occurring amino acids.
US Patent Application Publication No. 2003/0166510 teaches the use of ionic metal-peptide complexes in an amount effective to remodel the skin and diminish or remove skin blemishes. (Granted U.S. patents and published U.S. patent applications referenced herein are, to the extent pertinent, incorporated by reference.) Skin blemishes taught in this reference include scars (e.g., from wounds, acne), skin tags, calluses, benign skin moles, stretch marks, facial keratoses, solar lentigines or vitiligo spots. According to this reference, ionic metals—copper(II), tin(II), tin(IV), and zinc(II) and salts thereof—are complexed with chemically-synthesized di-, tri- and tetrapeptides. Phe-Phe and Gly-Gly are specifically taught as dipeptide fragments that may be complexed with the above-listed ionic metals.
Spanish Patent Application Publication No. ES2020148 teaches a biosurfactant consisting of a fatty acid chain of 9-17 carbon atoms, saturated or unsaturated, attached to the N-terminus of Arginine in any of the L-, D-, or DL forms followed by a second amino acid selected from any of the twenty naturally-occurring amino acids. The specific amino acid sequences Arg-Gly, Arg-Ser and Arg-Phe are taught.
Tripeptide-1, a synthetic peptide containing three amino acid residues—Glycine, Histidine and Lysine—is commercially-available from Vincience under the tradename Kollaren C.P.P. and as Kollaren by I.E.B. The mixture of Tripeptide-1 with water, urea, glucose and Guanidine HCl is sold as Kollaren by Atrium Biotechnologies. The Gly-His-Lys sequence is described in the literature as a scavenger of reactive carbonyl species (“RCS”) which are byproducts of cellular metabolic processes including lipid peroxidation and glycation. RCS have been associated with crosslinking of collagen and attendant loss of skin elasticity. See, Puig at al., “Peptides as Active Ingredients in Cosmetics,” Cosmetics and Toiletries Manufacture Worldwide, pp. 121-125. This amino acid sequence does not have a CMC. Moreover, it does not form polymeric aggregates. Accordingly, it is not a biosurfactant within the scope of the present invention.
Acetyl Tripeptide-1 is the reaction product of acetic acid and is therefore not a biosurfactant within the scope of the present invention. As a general matter, acetylation does not confer sufficient amphipathic properties needed to function as biosurfactant.
Biotinyl tripeptide is formed by grafting vitamin H (biotin) on the tripeptide Gly-His-Lys. US Patent Application Publication No. 2006/0067905 describes a method for treating hair loss by administering oleanolic acid, apigenin and Biotinyl-Gly-His-Lys. The biotinyl moiety does not confer sufficient hydrophobicity to produce biosurfactant properties.
Palmitoyl Tripeptide-1, also described as Pal-GKH, is the reaction product of Tripeptide-1 and palmitic acid. It is available from Sederma under the tradename Lipo-GKH. When acid-terminated, this lipo-oligopeptide sequence has no measurable antimicrobial activity; moreover, it is toxic to mammalian cells at comparatively low concentrations (e.g., LD50 of about 50 ppm). For these reasons, this lipo-oligopeptide is not within the scope of the present invention.
US Patent Application Publication No. 2004/0120918 at Paragraph #0008 describes Pal-Gly-His-Lys as Biopeptide CL available from Sederma. The INCI name for this tripeptide is Palmitoyl Oligopeptide which, according to the INCI Dictionary, is the palmitic acid ester of a synthetic peptide of two or more of the following amino acid residues: Alanine, Arginine, Aspartic Acid, Glycine, Histidine, Lysine, Proline, Serine or Valine.
Pal-Gly-His-Lys with the free acid or amide at the C-terminus exhibit significant toxicity in mammalian cell lines (i.e., having LD50<100 in 37-year-old female fibroblast cells) and for this reason are not within the scope of the present invention.
US Patent Application Publication No. 2004/0132667 teaches a sequence of three amino acids—Glycine, Histidine and Lysine residues. A preferred tripeptide has the specific amino acid sequence Gly-His-Lys. Analogs of this sequence are taught to include those in which one or more of the three amino acids are reorganized or rearranged within the sequence (e.g., Gly-Lys-His). This publication also teaches substitution of up to two of the three amino acids. Amino acids that may be substituted for Gly are taught to have an aliphatic side chain such as, without limitation, beta-Ala, Ala, Val, Leu, Pro and Ile. Of these, Ala, Leu and Ile are preferred. Amino acids that are taught to be substituted for Lys or His include those having a side chain that includes, predominantly, a charged nitrogen at a pH of about 6 (e.g., Pro, Lys, Arg, His, Desmosine and Isodesmosine). Most preferably, Lys is replaced with Ornithine, Arginine, or Citrulline.
The '667 application further teaches attaching to the above-described substituted or rearranged amino acid sequences acyl-moieties derived from: acetic acid, capric acid, lauric acid, myristic acid, octanoic acid, palmitic acid, stearic acid, behenic acid, linoleic acid, linolenic acid, lipoic acid, oleic acid, isostearic acid, elaidoic acid, 2-ethylhexaneic acid, coconut oil fatty acid, tallow fatty acid, hardened tallow fatty acid, palm kernel oil fatty acid, lanolin fatty acid. These derivatives are further taught to be straight-chain or branched-chain, long or short chain, saturated or unsaturated, substituted with a hydroxy, amino, acyl amino, sulfate or sulfide groups, or unsubstituted. Preferred acyl groups are taught to include palmitoyl and myristoyl. By teaching replacement of Lysine with Alanine or Arginine, and acylating the resulting three amino acid sequence with a palmitoyl or myristoyl group, the '667 Publication teaches the following acylated peptides: Pal-Gly-His-Arg; Pal-Arg-His-Ala; Pal-Arg-His-Gly; Pal-Ala-His-Arg; Myr-Gly-His-Arg; Myr-Arg-His-Ala; Myr-Arg-His-Gly; Myr-Ala-His-Arg. This patent publication does not, however, teach C-terminus amidation for these acylated tripeptides or the antimicrobial, stimulatory and/or proliferative properties of the polymeric biosurfactants of the present invention. Moreover, the tripeptides disclosed in this publication have only one positively-charged amino acid residue at neutral pH. As discussed below, the polymeric biosurfactants of the present invention largely contain two, and often, three positively-charged amino acid residues.
Tripeptide-2 is a synthetic peptide available under the tradename I.E.L. from Vincience. According to the INCI Dictionary, it contains two amino acid residues—Tyrosine and Valine. This amino acid sequence does not have a CMC and is therefore not a biosurfactant within the scope of the present invention.
Tripeptide-3, having the amino acid sequence Gly-His-Arg, has been disclosed in marketing materials by Therapeutic Peptides Inc. This amino acid sequence does not have a CMC and is therefore not a biosurfactant within the scope of the present invention.
The scientific literature reports that Gly-His-Lys-Cu, a copper tripeptide, has a stimulatory effect on collagen synthesis by fibroblasts. See Maquart F X at al., FEBS Lett. 238(2):343-6 (1988). See also, Oddos T et al. “Requirement of Copper and Tripeptide Glycyl-L-Histidyl-L-Lysine-Cu (GHK) Complex Formation for Collagen Synthesis Activity in Normal Human Dermal Fibroblasts” presented at the 60th Annual Meeting American Academy of Dermatology (New Orleans, La., February 2002). Additionally, this copper tripeptide has been reported to promote wound healing. See, Fish, et al., Wounds 3:171 (1991); Mulder et al., Wound Rep. and Regen. 2: 259 (1994).
Cosmetic use of Gly-His-Lys-Cu is described in U.S. Pat. Nos. 5,135,913 and 5,348,943 both assigned to ProCyte Corporation. Commercially, this copper tripeptide is used as an ingredient in Neutrogena Visibly Firm Night Cream as well as in products offered by ProCyte Corp. under the brand names Simple Solutions® Anti-Aging Skin Care (sold through spas and aestheticians) and Neova® (sold through dermatologists).
German Patent Application DE 41 27 790 A1, published on Feb. 25, 1993, teaches the use of Mg, Mn, Zn and Ge complexes of Gly-His-Lys to improve the condition of the skin. The Bibliographic Data for this patent application, as published on the European Patent Office website espacenet.net, also teaches tripeptides where each of the three constituent amino acids of the peptide is one of Lysine, Hydroxylysine, Proline, Hydroxyproline, Arginine, Glycine or Histidine. More particularly, a tripeptide conforming to the formula B1-B2-B3 is taught, where each of B1, B2 and B3 are one of the seven above-listed amino acids.
German Patent Application DE 42 44 418 A1 teaches cosmetic and pharmaceutical compositions containing Gly-His-Lys and Gly-Asp-Ser, both as tripeptides as well as part of a longer peptide moiety at concentrations of 1 picoM to 0.01M. These compositions are taught to be prepared either by mild hydrolysis of collagen, elastin, keratin or connective tissue with hydrochloric acid or partial hydrolysis using C. histolyticum collagenase. Among the disclosed anti-aging skin care applications are stimulation of collagen synthesis and scavenging of free radicals.
French Patent Application FR 2 826 577 A1 teaches peptides containing the sequence Lys-Pro-Val. According to this application, topical application of compositions containing this sequence increases the expression of genes coding for enzymes involved in the synthesis of epidermal lipids (i.e., cholesterol, fatty acids, and sphingolipids), thereby improving skin barrier function. The disclosed peptide sequence has no CMC and is therefore not a biosurfactant within the scope of the present invention.
U.S. Pat. No. 5,493,894 teaches compositions for treating skin wrinkles containing tri-, tetra- and pentapeptide moieties composed of at least three Arginine or Lysine residues. Among the specifically disclosed tripeptides are: (i) H-Arg-Lys-Arg-OH; (ii) H3C—C(O)-Arg-Lys-Arg-NH2. These two specifically-disclosed sequences do not have CMCs and therefore are not biosurfactants within the scope of the present invention.
US Patent Application Publication No. 2003/0166510 teaches the use of ionic metal-tripeptide complexes in which one of copper(II), tin(II), tin(IV), or zinc(II) is complexed with the following amino acid sequences: Gly-His-Lys; Gly-Gly-His; His-Gly-Gly; Gly-Gly-Gly; Ala-Gly-His; Gly-Cys-Gly; His-Gly-His. The above described metal-tripeptide complexes do not have CMCs and therefore are not biosurfactants within the scope of the present invention.
US Patent Application Publication 2006/0013794 teaches cosmetic, dermatological and/or pharmaceutical compositions comprising tri-, tetra-, penta-, hexa-, hepta- and nonpeptides containing the amino acid sequence Arg-Gly-Ser.
Tetrapeptide-1 is the INCI name assigned to a synthetic peptide containing four amino acid residues—Leucine, Proline, Threonine and Valine. It is sold under the tradename I.E.L. Leuococytar Elastase Inhibitor by Vincience. As discussed above, amino acid sequences alone do not have sufficient hydrophobicity to self-aggregate and, therefore do not have a CMC and are not biosurfactants within the scope of this patent. Moreover, none of the four amino acids in this compound have a charge. For this additional reason, this compound is not within the scope of the present invention.
Tetrapeptide-4 is the INCI name assigned to a synthetic tetrapeptide sold under the tradename Collasyn 4 GG by Therapeutic Peptides Inc. It contains three amino acid residues—Glycine, Glutamic Acid and Proline. More particularly, this peptide has the sequence Gly-Glu-Pro-Gly. For the reasons discussed above, this amino acid sequence is not a biosurfactant (i.e., no CMC, no self-aggregation) within the scope of the present invention.
Therapeutic Peptides Inc. has also disclosed the acylated amino acid sequence Myr-Gly-Glu-Pro-Gly under the tradename Collasyn 414 GG. At concentrations of 500 ppm or less, this sequence does not inhibit the growth of E. coli, P. acnes, P. aeruginosa, S. aureus and/or C. albicans and accordingly is not within the scope of the present invention.
Acetyl Tetrapeptide-1 is the reaction product of acetic acid and a synthetic peptide containing three amino acid residues—Glycine, Histidine and Lysine. It is sold under the tradename Kollaren 6 by I.E.B. For the reasons discussed above, this amino acid sequence is not a biosurfactant (i.e., no CMC, no self-aggregation) within the scope of the present invention.
Acetyl Tetrapeptide-2 is the reaction product of acetic acid and a synthetic peptide containing four amino acid residues—Aspartic Acid, Lysine, Tyrosine and Valine. Manufactured by I.E.B., the product is sold under the tradename Thymulen 4 by Atrium Biotechnologies. Product literature describes Thymulen 4 as a biomimetic peptide derived from thymopoietin having skin regenerative properties. For the reasons discussed above, this amino acid sequence is not a biosurfactant (i.e., no CMC, no self-aggregation) within the scope of the present invention.
Rigin, a tetrapeptide having the sequence Gly-Gln-Pro-Arg, is reported in the scientific literature by Veretennikova, et al., Int. J. Peptide Protein Res., 17:430 (1981). Palmitoyl Tetrapeptide is described in the INCI Dictionary as the reaction production of palmitic acid and a synthetic peptide containing Glycine, Glutamine, Proline and Arginine. It is commercially-available from Sederma.
At concentrations of 500 ppm or less, the acylated amino acid sequence Pal-Gly-Gln-Pro-Arg-acid does not inhibit the growth of microorganisms including E. coli and P. aeruginosa, and accordingly is not within the scope of the present invention. Moreover, both this compound and its amide-terminated analog exhibit significant toxicity in mammalian cell lines (i.e., having LD50<100 in 37 year-old female fibroblast cells).
Eyeliss is the tradename of a raw material concentrate combining two peptides and is marketed by Sederma for helping to reduce the appearance of puffiness and dark circles under the eyes. As described in International Patent Application PCT FR-03/00441, it is a combination of hesperidin methyl chalcone and two acylated peptide fragments—Valyl-Tryptophane and N-Palmitoyl-Gly-Gln-Pro-Arg. More generally, this PCT Application describes tri-, tetra- and pentapeptides beginning with a C2-C22 carbon chain and terminating in the sequence Pro-Arg-OH. According to U.S. Pat. No. 6,974,799, the Val-Trp dipeptide has no significant collagen stimulating activity and its combination with the Gly-Gln-Pro-Arg tetrapeptide does not exhibit any enhancement in this property over the levels realized by the use of the tetrapeptide alone.
Matrixyl 3000 is the tradename for a combination of two acylated peptides, N-Palmitoyl-Gly-Gln-Pro-Arg and N-Palmitoyl-Gly-His-Lys. U.S. Pat. No. 6,974,799 teaches topical compositions comprising (i) between about 0.00001% and about 0.5% (based on the total weight of the composition) of at least one “rigin-based tetrapeptide” (defined as Gly-Gln-Pro-Arg) and between about 0.00001% and about 1.0% of at least one tripeptide Gly-His-Lys, where the tripeptide is present in an amount greater than the tetrapeptide and (ii) at least one additional skin care ingredient. The disclosed composition is taught to be useful in reducing visible signs of aging and stretch marks as well as visible dark circles under the eyes.
DE 41 27 790 teaches the following tetrapeptides as part of an oligopeptide metal complex with Mg, Mn, Cu, Zn, Ge, Ni, Fe, Mo and Co: (i) Gly-His-Lys-Lys; (ii) Gly-His-Lys-Gly; (iii) Gly-His-His-Gly; (iv) Gly-His-His-Lys; (v) Gly-His-Arg-Lys; (vi) Gly-His-Arg-Gly; (vii) Gly-His-pro-Lys; Gly-His-Pro-Lys; (ix) Hyp-Gly-Lys-Lys; (x) Hyp-Gly-His-Lys; (xi) Hyp-Gly-Arg-Lys; (xii) Hyp-Gly-Pro-Lys; (xiii) Arg-Gly-Lys-Lys; (xiv) Arg-Gly-Lys-Lys; (xv) Arg-Gly-His-Lys; (xvi) Arg-Gly-His-Lys; (xvii) Arg-Gly-Arg-Lys; (xviii) Arg-Gly-Arg-Lys; (xix) Arg-Gly-Pro-Lys; and (xx) Arg-Gly-Arg-Lys, where Hyp is hydroxyproline.
The Bibliographic Data for German Patent Application DE 41 27 790, as published on ep.espacenet.com, also teaches tetrapeptides where each of the first three amino acids of the peptide is one of Lysine, Hydroxylysine, Proline, Hydroxyproline, Arginine, Glycine or Histidine and the fourth amino acid is the same as one of the preceding three amino acids. More particularly, tetrapeptides conforming to the formulae B1-B2-B3-B1, B1-B2-B3-B2 and B1-B2-B3-B3. The oligopeptide metal complexes as disclosed in this application do not have a CMC and do not self-aggregate. For these reasons they are not biosurfactants within the scope of the present invention.
U.S. Pat. No. 5,493,894 specifically teaches compositions for treating skin wrinkles containing the following tetrapeptides: (i) H-Arg-Gly-Arg-Lys-OH and (ii) H-Lys-Arg-Ser-Arg-NH2. These are not biosurfactants and thus are not within the scope of the present invention.
US Patent Application Publication No. 2003/0166510 teaches ionic metals complexed with the tetrapeptide Gly-His-Lys-His. Topical compositions comprising this metal ion/tetrapeptide complex are taught to be useful in diminishing or removing skin blemishes.
Therapeutic Peptides Inc. has disclosed in trade literature the amide-terminated VPAA tetrapeptide sequence as well as Myristoyl Tetrapeptide-5, an acylated synthetic peptide having the VPAA sequence. The latter is commercially available under the tradename Collasyn 414 VA. At concentrations of 500 ppm or less, this acylated amino acid sequence does not inhibit the growth of microorganisms, including E. coli. Moreover, this sequence contains no charged amino acid residues. Accordingly, for these reasons, Myr-Val-Pro-Ala-Ala is not within the scope of the present invention.
U.S. Pat. No. 4,665,053 teaches “bifunctional” synthetic lipopeptides, which are further defined as functioning both as inhibitors of elastolytic activity and protectors of elastic fibers. Additionally, these lipopeptide moieties are described as being capable not only of recognizing and becoming fixed on elastic fiber but also of recognizing and neutralizing the active site of elastases. More particularly, this reference teaches lipopeptides having in two sequential L-Alanine residues conforming to the formula: R—X—(P1)x-(L-Ala-L-Ala-P2)-A where P1 is an amino acid sequence, two to eight residues in length; x is 0 or 1; R is an acylated hydrophobic carboxylic acid. P2 is taught to be one of L-Ala, L-Val, L-Pro-L-Ala or L-Pro-L-Val. A is the C-terminus in the form of acid, aldehyde, alcohol, amide or chloromethyl ketone.
Palmitoyl Pentapeptide-2 is the reaction of palmitic acid and a synthetic peptide consisting of four amino acid residues—Tyrosine, Glycine, Phenylalanine and Leucine. It is available from Sederma. This acylated amino acid sequence contains no charged residues and accordingly is not within the scope of the present invention.
Pentapeptide-3 is sold under the tradename Matrixyl by Sederma. It described in the INCI Dictionary as the reaction product of palmitic acid and a synthetic peptide consisting of Lysine, Threonine and Serine residues. The INCI Dictionary does not list the amino acid sequence of this material. Without further information, and interpreting “consisting” to mean that only the three listed amino acid residues are present in the product, this reference would teach sixty combinations without suggesting which one(s) would have particular properties.
As disclosed in trade literature and marketing materials of finished goods companies, the amino acid sequence of the Matrixyl pentapeptide is Lys-Thr-Thr-Lys-Ser. This compound is further described in U.S. Pat. No. 6,620,419 which claims peptides according to the formula: R1—X-Thr-Thr-Lys-(AA)n-Y. X is defined as one of seven amino acids, with D or L orientation. Among the seven amino acids taught at the X position is Lysine. R1 is taught to be hydrogen or a fatty acid chain of 2 to 22 carbons, which includes palmitoyl. (AA)n is taught to represent a chain of n amino acids where n varies from 0 to 5. Y is defined as OR2 or NR2R3, where R2R2 may be hydrogen, resulting in acid and amide C-termini.
Pal-KTTKS-acid does not exhibit antimicrobial activity at a concentration of less than 500 ppm and therefore is not a polymeric biosurfactant within the scope of the present invention.
Collasyn 514KS is the tradename for Myristoyl Pentapeptide-3. This synthetic peptide contained Threonine, Serine and Lysine residues in the sequence Myr-KTTKS-amide and was available from Therapeutic Peptides Inc. This moiety does not result in an increase in soluble metabolic proteins, does not increase cell turnover, nor does it possess desired antimicrobial properties. For these reasons, Collasyn 514KS is not within the scope of the present invention.
DE 41 27 790 A1 teaches pentapeptides of the following sequences as being complexed with Mg, Mn, Cu, Zn, Ge, Ni, Fe, Mo and Co: (i) Hyp-Gly-Lys-Hyp-Gly; (ii) Hyp-Gly-His-Lys-Gly; (iii) Gly-Pro-Lys-Gly-Pro. These peptides are not acylated and therefore are not biosurfactants within the scope of the present invention.
The Bibliographic Data for German Patent Application DE 41 27 790, as published on ep.espacenet.com, also teaches pentapeptides where (i) each of the first three amino acids is one of Lysine, Hydroxylysine, Proline, Hydroxyproline, Arginine, Glycine or Histidine and (ii) the fourth and fifth amino acids are the same as one of the preceding three amino acids. More particularly, the bibliographic data teaches pentapeptides corresponding to the following six formulae: (i) B1-B2-B3-B1-B2; (ii) B1-B2-B3-B2-B3; (iii) B1-B2-B3-B2-B3; (iii) B1-B2-B3-B2-B1-B3; (iv) B1-B2-B3-B2-B1; (v) B1-B2-B3-B3-B2; and (vi) B1-B2-B3-B3-B1, where each of B1, B2 and B3 is Lysine, Hydroxylysine, Proline, Hydroxyproline, Arginine, Glycine or Histidine. These peptides are not acylated and therefore are not biosurfactants within the scope of the present invention.
Acetyl Pentapeptide-1 is sold under the tradename Thymulen by Atrium Biotechnologies. It is the reaction product of acetic acid and Pentapeptide-1. In product literature, Thymulen is described as inducing the secretion granulocyte-macrophage colony stimulating factor, resulting in a multiplication and a differentiation of keratinocytes. This peptide moiety does not have a measurable CMC and is therefore not a biosurfactant within the scope of the present invention.
Therapeutic Peptides Inc. has also offered for sale the following amide-terminated amino acid sequences: EVEDQ; DSDPR; GRKGD; GEESN; KKALK; KRGDR; LPPSR. Because these peptide moieties are not acylated they do not have a CMC and are not biosurfactants within the scope of the present invention.
U.S. Pat. No. 6,492,326 claims pentapeptides and/or pentapeptide derivatives and mixtures thereof in combination with an “additional skin care active” in a dermatologically-acceptable carrier. These additional skin care actives are taught to include di-, tri-, and tetrapeptides (and their derivatives) as well as retinoids, hydroxy-acids, anti-inflammatory, anti-fungal and anti-microbial agents.
WO97/18235 entitled “Peptide Conjugates, Use Thereof as a Drug and Compositions Containing Same” published in May 1997. Pentapeptides and pentapeptide derivatives are disclosed at page 6 #s 2, 5, 7, 9, 11 and at page 7 #14. Additional skin care actives, specifically antifungal and antimicrobial agents are taught in combination with the disclosed peptides at page 8, lines 19-24. WO97/18235 also teaches that the disclosed peptides and their derivatives can be used in creams, gels, milks, lotions, and sprays with excipients well-known in the cosmetics industry. Page 3 of this application further teaches that the peptide sequences can be acylated with straight-chain or branched, saturated or unsaturated, C1-C20 monocarboxylic acids. More particularly, this application discloses lipo-oligopeptides having the specific amino acid sequence Gly-His-Lys within the oligopeptide. The biosurfactants of the present invention do not contain this specific sequence.
Hexapeptides of the following amino acid sequences are taught by DE 41 27 790 A1 as being complexed with one of Mg, Mn, Cu, Zn, Ge, Ni, Fe, Mo and Co: (i) Gly-Pro-Arg-Gly-Pro-Hyp; (ii) Gly-His-Hyp-Gly-Lys-Pro; (iii) Gly-Lys-Pro-Gly-Arg-Hyp; (iv) Gly-Pro-Hyp-Gly-Pro-Pro; (v) Gly-His-Arg-Gly-His-Lys. Because these peptide moieties are not acylated they do not have a CMC and are not biosurfactants within the scope of the present invention.
Hexapeptide-1 is a synthetic peptide consisting of six amino acid residues—Alanine, Arginine, Histidine, Leucine, Phenylalanine and Tryptophan. Because these peptide moieties are not acylated they do not have a CMC and are not biosurfactants within the scope of the present invention.
Acetyl Hexapeptide-1, the reaction product of acetic acid and Hexapeptide-1, is sold under the tradename Melitane by Vincience. This peptide moiety does not have a measurable CMC and is therefore not a biosurfactant within the scope of the present invention.
Melitane 5 PP is the tradename for dextran and Acetyl Hexapeptide-1. Melitane 5 PS is the tradename for water dextran and Acetyl Hexapeptide-1. The PP and PS designators indicate, respectively, peptide powder and peptide solution. The PS product is described in trade literature as a peptide that mimics the activity of alpha melanocyte stimulating hormone, stimulating melanogenesis. Both are commercially available from I.E.B. This peptide moiety does not have a measurable CMC and is therefore not a biosurfactant within the scope of the present invention.
Acetyl Hexapeptide-3 is a synthetic peptide consisting of three amino acids—Arginine, Methionine and acetylated Glutamic Acid. It is sold under the tradename Argireline by Lipotec. This peptide moiety does not have a measurable CMC and is therefore not a biosurfactant within the scope of the present invention.
Hexapeptide-4 is a synthetic peptide containing Lysine, Threonine and Serine residues and is commercially available from Therapeutic Peptides Inc. under the tradename Collasyn 6KS. This peptide moiety is not acylated and does not have a CMC; accordingly, it is not a biosurfactant within the scope of the present invention.
Hexapeptide-5 is a synthetic peptide containing Valine, Tyrosine, Glutamic Acid, Proline and Isoleucine residues. It is commercially available from Therapeutic Peptides Inc. under the tradename Collasyn 6VY. This peptide moiety is not acylated and does not have a CMC; accordingly, it is not a biosurfactant within the scope of the present invention.
Hexapeptide-6 is a synthetic peptide having the sequence VEPIPY. It is commercially-available from Therapeutic Peptides, Inc. This peptide moiety is not acylated and does not have a CMC; accordingly, it is not a biosurfactant within the scope of the present invention.
The scientific literature describes the chemotactic activity of several components of the extracellular matrix including collagen, fibronectin, elastin and tropoelastin, the soluble precursor of elastin. The chemotactic activity of elastin for fibroblasts has reported to be associated with the repeating hexapeptide sequence Val-Gly-Val-Ala-Pro-Gly. See Senior R M et al., J. Cell Biol. 99(3): 870-874 (1984). As a sequence, Val-Gly-Val-Ala-Pro-Gly has also been reported to stimulate the growth of human skin fibroblasts. See, Kamoun A. et al., Cell Adhes. Commun., 3(4): pp. 273-81 (1995). Because this peptide sequence is not acylated it does not have a CMC and is not a biosurfactant within the scope of the present invention.
Palmitoyl-Val-Gly-Val-Ala-Pro-Gly is an acylated hexapeptide available from Sederma under the tradename Biopeptide EL. The INCI name for Biopeptide EL is Palmitoyl Oligopeptide. Because this peptide sequence does not contain at least one charged amino acid moiety it is not within the scope of the present invention.
US Patent Application Publication No. 2004/0120918 teaches the use of a ceramide to improve the anti-aging activity of a polypeptide (or an acylated polypeptide) having an amino acid sequence of from 3 to 12 amino acids in length. The '918 Publication teaches N-acyl derivatives of the Val-Gly-Val-Ala-Pro-Gly hexapeptide, where the acyl chain is an alkoyl of 2-22 carbons, linear or branched, saturated or unsaturated, hydroxylated or non-hydroxylated. This publication more specifically teaches Biopeptide EL in combination with three ceramides, n-stearoyl-dihydrosphingosine, trihydroxypalmitamidohydroxy-propylmyristyl ether or palmitamidomyristylserinate. This specific hexapeptide amino acid sequence is not contained within the polymeric biosurfactants of the present invention.
The commercial product Bio-Bustyl, available from Sederma is a combination of Pal-VGVAPG and Pal-GHK. The INCI name for this compound is Glyceryl Polymethacrylate-Rahnella/Soy Protein Ferment-Water (Aqua)-Propylene Glycol-Glycerin-PEG-8-Palmitoyl Oligopeptide. Pal-VGVAPG does not contain a charged amino acid residue and is therefore not a polymeric biosurfactant within the scope of the present invention.
Myristoyl Hexapeptide-6 is available from Therapeutic Peptides Inc. under the tradename Collasyn 614VG. According to the INCI Dictionary, it is the reaction product of Myristic Acid with a synthetic peptide containing Valine, Glycine, Alanine and Proline residues. Collasyn 614VG does not contain a charged amino acid residue and is therefore not a polymeric biosurfactant within the scope of the present invention.
Acetyl Hexapeptide-7 is described in the INCI Dictionary as the reaction product of acetic acid and Hexapeptide-7. It is sold under the tradename Melitane 5 by Atrium Biotechnologies. Melitane 5 PP and 5 PS are mixtures of dextran and Acetyl-Hexapeptide-7, with 5 PS also containing water. Both are available from I.E.B. This peptide moiety does not have a measurable CMC and is therefore not a biosurfactant within the scope of the present invention.
Palmitoyl Oligopeptide is also the INCI designation for Biopeptide EN from Sederma. As further described in US Patent Application Publication No. 2002/0025303, Biopeptide FN is composed of Arginine, Aspartic Acid, Glycine, and Serine. None of the compositions of the present invention are composed of all four of these amino acids.
Hexapeptide-8 has been sold by Therapeutic Peptides Inc. as the amide-terminated sequence Ser-Thr-Lys-Thr-Thr-Lys. Because this peptide moiety is not acylated it does not have a CMC and is not a biosurfactant within the scope of the present invention.
WO 9962482 describes alkyl-heptapeptides where a 9- to 13-membered carbon chain is bound to the N-terminus of the heptapeptide sequence Glu-Leu-Leu-Val-Asp-Leu-X1, where X1 is an amino acid selected from the group consisting of the twenty naturally-occurring amino acids, hydroxyproline and homoserine. None of the compositions of the present invention contain the six amino acid sequence Glu-Leu-Leu-Val-Asp-Leu.
Nonapeptide-1 in a mixture with water and dextran is sold under the Melanostatine 5 by Atrium Biotechnologies. According to its product literature, it is a biomimetic peptide for skin whitening. Because this peptide moiety is not acylated it does not have a CMC and is not a biosurfactant within the scope of the present invention.
Therapeutic Peptides Inc. has offered for sale an amide-terminated nonapeptide having the sequence VQGEESNDK. Because this peptide moiety is not acylated it does not have a CMC and is not a biosurfactant within the scope of the present invention.
US Patent Application Publication 2005/0124545 teaches cosmetic, dermatological and/or pharmaceutical compositions containing fifteen amino acid sequence X1-Y-Phe-Thr-X2-Ala-Thr-Z-Ile-X3-Leu-X4-Phe-Leu-X5. Each of X1, X2, X3, X4 and X5 is defined as one of Arg, Lys or His. Y is defined as either Asp or Glu. Z is defined as either Asn or Gln. These specific amino acid sequences are not contained within the polymeric biosurfactants of the present invention. Moreover, because the peptide moieties according to the above formula are not acylated they do not have a CMC and are not biosurfactants within the scope of the present invention.
U.S. Pat. No. 5,492,894 to Bascom et al. teaches peptides having three to six amino acid residues, two of which are Arg and one of which is Lys, useful in the cosmetic treatment of mammalian skin wrinkles. The following thirty-eight amino acid sequences are disclosed: (1) RGRK; (2) KRSR; (3) RSRK; (4) YRSRKY; (5) YRSRK; (6) RSRKY; (7) TYRSRKYS; (8) SYRSRKYT; (9) SYRSRKYS; (10) TYRSRKYT; (11) RSRKYT; (12) TYRSRK; (13) RSR KYS; (14) SYRSRK; (15) YRSRKYT; (16) TYRSRKY; (17) YRSRKYS; (18) SYRSRKY; (19) NTYRSRKYSS; (20) NSYRSRKYTS; (21) NSYRSRKYSS; (22) NTYRSRKYTS; (23) RSRKYTS; (24) NTYRSRK; (25) RSRKYSS; (26) NSYRSRK; (27) YRSRKYTS; (28) NTYRSRKY; (29) YRSRKYSS; (30) NSYRSRKY; (31) TYRSRKYSS; (32) NTYRSRKYS; (33) SYRSRKYTS; (34) NSYRSRKYT; (35) SYRSRKYSS; (36) NSYRSRKYS; (37) TYRSRKYTS; and (38) NTYRSRKYT. Alkyloyl is among the groups that may be attached to the first amino acid in the sequence.
U.S. Pat. No. 6,875,744 teaches non-acylated peptides sequences, five to twenty-two amino acids in length comprising at least 80% Phenylalanine, Leucine, Alanine, and Lysine residues. The peptides disclosed in the '744 patent do not have CMCs and are not biosurfactants within the scope of the present invention.